Nuclear specific glycoprotein representative of a unique pattern of glycosylation

Journal Article · Sun Jan 25 04:00:00 EST 1987 · J. Biol. Chem.; (United States)

OSTI ID:6943076

Schindler, M; Hogan, M; Miller, R; DeGaetano, D

Whole rat liver nuclei were reacted with UDP-(/sup 14/C)galactose in the presence of bovine beta(1----4) galactosyltransferase. The reaction mixture was electrophoresed on a reducing sodium dodecyl sulfate-polyacrylamide gel. Autoradiograms of the gel demonstrated a major labeled broad band migrating with an apparent molecular weight of 65,000-66,000. A number of other less prominently labeled bands were also present. The labeled 65,000-66,000 band when cut from the gel and subjected to alkaline reduction while in the gel matrix exclusively yielded a /sup 14/C-labeled disaccharide that co-migrated with a (/sup 14/C)Gal-GlcNAcol standard in descending paper chromatography. Treatment of this disaccharide with beta-galactosidase (beta-D-galactoside galactohydrolase; EC 3.2.1.23) from Aspergillus niger removed all the (/sup 14/C)galactose label. Treatment of the labeled 65,000-66,000 polypeptide with Endoglycosidase F, however, did not remove the (/sup 14/C)galactose label. Western transfer blots of sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels performed with horseradish peroxidase-labeled succinyl wheat germ agglutinin, a lectin specific for GlcNAc, on unlabeled nuclei revealed a dominant band at 63,000-64,000. Subjecting /sup 14/C-labeled nuclei to this procedure resulted in a shift of the major horseradish peroxidase-labeled succinyl wheat germ agglutinin band to 65,000-66,000. The shifted band was coincident with the (/sup 14/C)galactose band as visualized on an autoradiogram. A survey of other rat tissue nuclei revealed the same spectrum of (/sup 14/C)galactose acceptor proteins with a dominant 65,000-66,000 galactose-labeled band.

• Research Organization: Michigan State Univ., East Lansing
• OSTI ID: 6943076
• Journal Information: J. Biol. Chem.; (United States), Journal Name: J. Biol. Chem.; (United States) Vol. 3; ISSN JBCHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
ANIMALS
AUTORADIOGRAPHY
BODY
CARBOHYDRATES
CARBON 14 COMPOUNDS
CELL CONSTITUENTS
CELL NUCLEI
CHROMATOGRAPHY
DIGESTIVE SYSTEM
ELECTROPHORESIS
ENZYMES
GALACTOSE
GLANDS
GLUCOPROTEINS
HEXOSES
LABELLED COMPOUNDS
LIVER
MAMMALS
MOLECULAR WEIGHT
MONOSACCHARIDES
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PROTEINS
RATS
RODENTS
SACCHARIDES
SEPARATION PROCESSES
TRANSFERASES
VERTEBRATES